Postlab of Frog

Experiments
 Exercise 11
Pithing the Frog
PITHING
 make the animal free from pain
RA 8485 Animal Welfare Act
 destroy the Nervous System
 pithing/dissecting needle
 foramen magnum
- cup-like depression
- located between the skull and the atlas (first
cervical vertebra)
 single pithing – brain is destroyed
 double pithing – brain and spinal cord are
destroyed
 Signs of Successful Pithing
• Frog will remain limp
• Legs are extended forward or
outstretched
and frog will quiver

 Single pithing will only destroy

the brain. Frog will still be able to:
 breath
 muscles will still contract
 heart will still beat
 Proper way of restraining the frog
during pithing:
• Legs should be outstretched and held
tightly between ring and small finger
• Head bent forward , pressure exerted on
top of the head by index finger
• Head is bent over the third finger
 Exercise 12
Muscle-Nerve Preparation
 Parts of Muscle-Nerve Preparation
• Sciatic nerve – whitish thread-like
structure lying in between
the thigh muscle close to the
femur
• Femur
• Thigh muscles
• Gastrocnemius – large muscle at the postero- inferior
part of the lower leg opposite the
peroneus
• Tendon of Achilles

 remove peroneus, shinbone (tibiofibula) and foot (pes)

 Muscle- nerve preparation is placed in amphibian Ringer’s
solution
 Exercise 13
Application of Different Forms of
Stimuli
Stimulus
- any change in the environment
- Tissues maybe stimulated by:
a. mechanical stimulus (tie and pinch the sciatic
nerve)
b. thermal stimulus ( hot and cold water)
c. chemical stimulus (NaCl )
d. electrical stimulus – (stimulator) - preferred
1. intensity and duration can be controlled
2. it maybe applied and removed easily
3. changes that the electrical stimulus
produces is reversible and do not
damage the tissues
 Stimulus Applied Response

1. Mechanical a. Tying single

b. Pinching single
2. Thermal a. Hot water single

b. Cold single
water
3. Chemical (NaCl) series

4. Electrical a. Direct single

 ANSWER TO QUESTIONS
1. Which among the different forms of stimuli
applied gives a better result?
Ans: Among the different forms of stimuli applied,
electrical stimulus gave the better result
1. intensity and duration can be controlled
2. it maybe applied and removed easily
3. change that the electrical stimulus
produces is reversible and do not
damage the tissues
2. Why is the chemical stimulus not frequently
applied in the laboratory?

Ans: Chemical stimulus is not frequently use in

the laboratory because it damages the tissue.
Threshold stimulus
- stimulus whose strength is just enough to
elicit a response
Subminimal stimulus
- weaker than threshold stimulus
- does not cause any response
Maximal Stimulus
- type of stimulus that causes the greatest response
Supramaximal stimulus
- strength is greater than maximal stimulus but
response is similar to that of maximal stimulus
 Exercise 14
The Spinal Frog
• Spinal frog- single pithing
• Normal frog ( only the semicircular canal is
destroyed)
 Condition Spinal Frog Normal Frog
1. Position of :
a. head Drooping Erect
b. eyes Half closed Wide open
c. limbs Extended Contracted

2. Pinching the toes No response

3. Pinching the skin No response
4. Pinching the hind toes No response
5. Response on the other No response
toes
6. Position of the body Tilted to one Normal position
side (not
balanced
7. Evidence of sight No response Frog moved

8. Evidence of No response frog moved

hearing

9. Touching the No response Eyes closed

cornea

10. Destruction of Not balanced Normal position

semicircular canal
 Exercise 15
Complex Coordinated and
Uncoordinated Reflexes
 Condition Observation Type of Reflex

Before injection of
strychnine
a. concentrated Frog flexed the leg  all are complex
HNO3 coordinated
b. immersion in withdraws the leg reflexes (orderly
hot water sequence of
muscular
contractions
After injection of
strychnine (dorsal
lymph sac)
a. concentrated Stiffening of the  Uncoordinated
HNO3 leg (convulsive reflexes
b. immersion in movements/spasm
hot water odic contractions)
 Answer to Questions
1. What type of reflexes was produced upon the first
application of acid?

Ans: The type of reflex produced upon the first

application of acid is complex coordinated.

After injection of strychnine and application of acid?

Ans: The strychnine solution causes convulsive

movements or spasmodic contraction because
strychnine binds with glycine which is a
neurotransmitter for relaxation and contraction
equilibrium therefore the muscle remains contracted.
3. Under what group of reflexes will you classify
the withdrawal reflex?

Ans: Withdrawal reflexes are classified under

complex coordinated reflex.
 Exercise 35
The Effect of Temperature on
Heart Rate
 Condition Results
heartbeats/minute

1. Intact heart 60-100 beats/min

2. heart in Ringer’s Slightly lower than intact
solution (room heart
temperature)

3. heart in warm  the higher the temp.

Ringer’s solution the higher the cardiac
rate
4. Heart in cold Ringer’s  the lower the temp.
solution the lower the cardiac
rate
 Sympathetic Parasympathetic

Force increases decreases

Rate increases decreases

Rhythm increases decreases

 Answer to Questions
1. How do increased calcium and potassium levels affect the
heart rate?

Ans: Increase calcium levels – increase heart rate

Increase potassium levels – lower heart rate

2. Differentiate sympathetic/parasympathetic stimulation

Ans: Sympathetic stimulation – increases cardiac

activities
Parasympathetic stimulation – decreases
cardiac activities
 Exercise 10
Isotonic and Isometric Contractions
 Isotonic Isometric
Contraction Contraction

Muscle length Shortens Muscle retains

original length
Muscle tension Less tension More tension

Inertia Greater Lesser

Force Lesser force Greater Force

Isotonic Contraction
- Muscle length changes but the force produced
does not

Isometric Contraction
- Is the condition in which muscle length does
not change regardless of the amount of force
generated by the muscle
 Answer to Questions:
1. Isotonic contractions
ex. Walking, writing, running
Isometric contractions
ex. Simply standing, pushing against a wall
2. Isotonic contraction
- muscle shortens, mechanical work
Isometric contraction
- muscle tightening but no shortening of
muscle
 Exercise 8
Simple Muscle Contraction
• Single pithing
• Muscle preparation (same as muscle nerve
preparation)
• Parts of muscle preparation?
• Parts removed?
• What muscle is used?

 to produce simple muscle contraction, electrical

stimulus is applied to the muscle.
Muscle is allowed to relax before application of
second stimulus
Myogram
- record of muscle contraction
Muscle twitch/Simple Muscle Contraction
- brief contraction of a muscle fiber in a motor
unit in response to a single action potential in
a motor neuron
Three Phases of Simple Muscle
Contraction
A-B lag or latent phase
- period after stimulation
- contraction is not
apparent
B-C contraction phase
- period when muscle fibers
contract (muscle fibers shorten)
C-D relaxation phase
- period when the myogram falls
- muscle returns to its resting state
 Exercise 9
Summation
1. Summation
- adding together of individual muscle
twitches to make a strong coordinated
movements
- brought about by sending at least 2
successive stimuli
- second stimulus comes before the end of
contraction or at the beginning of relaxation
period.
A - first baseline
B - first peak/second baseline
C - second peak

AB first contraction
BC second contraction
CD relaxation
2. Second contraction is higher
than the first contraction because
of the ff. reasons:
a. greater tension is a - first baseline
produced on the muscle b - first peak/second baseline
upon sending more than c - second peak

one stimulus.
b. second contraction is
higher than the first
because it starts at a
higher baseline
c. greater amount of Ca on the
second contraction
The End